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Updated: Aug 10, 2026

Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction
Published on: July 12, 2022
The use of nuclear and mitochondrial single nucleotide polymorphisms to identify cryptic species
N M Belfiore1, F G Hoffman, R J Baker
1Department of Forestry & Natural Resources, Purdue University, 195 Marstellar St, West Lafayette, IN 47907-2033, USA. nmb@uclink.berkeley.edu
Abstract:
There is growing interest in the use of single nucleotide polymorphisms for evolutionary and population genetics. We tested the efficacy of one of the available single nucleotide polymorphism techniques, single-base extension, in distinguishing four cryptic species of Microtus. Sequence data were available for these species at nuclear and mitochondrial loci and their identity could be independently confirmed using karyotypes. We found that the development and optimization of single nucleotide polymorphisms required extensive effort, and that the method accurately identified the correct nucleotide at single nucleotide polymorphism sites approximately 90% of the time at the conserved nuclear locus. Correct identification rates were much lower at the highly variable mitochondrial locus.
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